Core--Integrated Response to Toxic Perturbation

在技术进步和研究者需求的推动下，该核心已经从最初的提供DNA微阵列服务的目标扩展到包括蛋白质组学和代谢组学，从而提供了一种评估细胞和组织对毒性暴露反应的综合方法。首先，微阵列服务将为毒性终点的调查提供转录组学分析。为此，它将制备具有高密度阵列的载玻片（例如，20,000 元件），其代表来自人、小鼠或大鼠的转录基因。它还将根据需要制备载玻片，载玻片上有较小的阵列或（视情况而定）代表其他物种转录本或基因的DNA。Core将根据要求处理切片或培训实验室人员，并与Core B合作，协助数据处理。第二，核心将提供蛋白质组学服务，包括分析（蛋白质 分离和质谱鉴定）和分析翻译后修饰（磷酸化、巯基氧化）。核心还将提供代谢组学研究服务，如代谢子域分析和代谢指纹图谱（包括II期代谢）。代谢子域的验证分析包括对调节性氧化脂质（用于研究组织对炎症损伤和一般修复机制的反应）和结构/能量脂质（用于研究毒性对细胞/生物体能量利用的影响）的定量评估。代谢指纹图谱为广泛探索毒物对组织、细胞和生物液体小分子环境的影响提供了工具。整合的主题是找到对给定药物的特征性反应，识别早期反应的生物标志物，并找到最敏感的生物标志物的无效应水平。重点将是提高项目生产力和资源利用效率。因此，训练有素的核心人员将帮助规划实验，将集中精力从事技术要求高的职能，并在可行的情况下培训项目人员进行自己的实验。核心将提供数据储存和数据分析方面的协助。

Driven by technological advances and investigator demand, this Core has expanded from the original goal of providing DNA microarray service to include proteomics and metabolomics, thus providing an integrated approach to assessing cell and tissue responses to toxic exposure. First, the microarray service will provide transcriptomic profiling for investigation of toxic endpoints. To this end, it will prepare glass slides with high density arrays (e.g., 20,000 elements) of long oligonucleotides representing transcribed genes from human, mouse or rat. It will also prepare slides on demand containing smaller arrays or (subject to availability) DNA representing transcripts or genes from other species. The Core will process the slides or train lab personnel in this function as requested and, in concert with Core B, will assist in data processing. Second, the Core will provide proteomics services including profiling (with protein separation and mass spectrometric identification) and analysis of posttranslational modifications (phosphorylation, sulfhydryl oxidation). The Core will also provide services for metabolomic investigations such as profiling of metabolic subdomains and metabolic fingerprinting (including phase II metabolism). Validated analyses of metabolic subdomains include the quantitative assessment of both regulatory oxylipids (for the study of tissue responses to inflammatory injury and general repair mechanisms) and structural/energetic lipids (for the study of toxicant effects on cell/organism energy utilization). Metabolic fingerprinting provides tools for an expansive exploration of toxicant effects on the small molecule milieu of tissues, cells, and biofluids. Integrating themes are to find characteristic responses to given agents, identifying biomarkers of early response, and finding no effect levels for the most sensitive biomarkers of effect. The emphasis will be on increasing Project productivity and efficiency of resource utilization. Thus highly trained Core personnel will help in planning experiments, will concentrate their efforts on technically demanding functions, and will train Project personnel in conducting their own experiments where feasible. The Core will provide data storage and assistance in data analysis.